High-efficiency liquid chromatographic separation utilizing long monolithic silica capillary columns.
Long monolithic silica-C18 capillary columns of 100 microm i.d. were prepared, and the efficiency was examined using reversed-phase HPLC under a pressure of up to 47 MPa. At linear velocities of 1-2 mm/s, 100,000-500,000 theoretical plates could be generated with a single column (90-440 cm in length) using an acetonitrile-water (80/20) mobile phase with a column dead time (t0) of 5-40 min. It was possible to prepare columns with a minimum plate height of 8.5 +/- 0.5 microm and permeability of (1.45 +/- 0.09) x 10(-13) m(2). The chromatographic performance of a long octadecylsilylated monolithic silica capillary column was demonstrated by the high-efficiency separations of aromatic hydrocarbons, benzene derivatives, and a protein digest. The efficiency for a peptide was maintained for an injection of up to 0.5-2 ng. When three 100 microm i.d. columns were connected to form a 1130-1240 cm column system, 1,000,000 theoretical plates were generated for aromatic hydrocarbons with retention factors of up to 2.4 with a t0 of 150 min. The fact that very high efficiencies were obtained for the retained solutes suggests the practical utility of these long monolithic silica capillary columns.
Miyamoto
Kosuke
K
Department of Biomolecular Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
Hara
Takeshi
T
Kobayashi
Hiroshi
H
Morisaka
Hironobu
H
Tokuda
Daisuke
D
Horie
Kanta
K
Koduki
Kodai
K
Makino
Satoshi
S
Núñez
Oscar
O
Yang
Chun
C
Kawabe
Takefumi
T
Ikegami
Tohru
T
Takubo
Hirotaka
H
Ishihama
Yasushi
Y
Tanaka
Nobuo
N
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006841:Hydrocarbons, Aromatic / Q000302:isolation & purification; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism; D012822:Silicon Dioxide
D006841:Hydrocarbons, Aromatic; D010455:Peptides; D012822:Silicon Dioxide